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Chapter 1 From Classical Genetics to Molecular Genetics
1£®1 Introduction
1£®2 Classical genetics and the rules of trait inheritance
1£®2£®1 Gregor Mendel developed the formal rules of genetics
1£®2£®2 Mendel's laws have extensions and exceptions
1£®2£®3 Genes are arranged linearly on chromosomes and can be mapped
1£®2£®4 The nature of genes and how they determine phenotypes was long a mystery
1£®3 The great breakthrough to molecular genetics
1£®3£®1 Bacteria and bacteriophage exhibit genetic behavior and serve as model systems
1£®3£®2 Transformation and transduction allow transfer of genetic information
1£®3£®3 The Watson-Crick model of DNA structure provided the final key to molecular genetics
Chapter 2 Structures and Functions of Proteins and Nucleic Acids
2£®1 Introduction
2£®2 Proteins
2£®2£®1 Amino acids and peptides
2£®2£®2 Levels of structure in the polypeptide chain
2£®2£®3 Protein folding
2£®2£®4 Protein destruction
2£®3 Nucleic acids
2£®3£®1 Chemical structure of nucleotides
2£®3£®2 Physical structure of DNA
2£®3£®3 Physical structures of RNA
2£®3£®4 One-way flow of genetic information
2£®3£®5 Methods used to study nucleic acids
Chapter 3 Recombinant DNA£º Principles and Applications
3£®1 Introduction of homologous recombination and cloning
3£®1£®1 The beginnings of recombinant DNA technology
3£®1£®2 Homologous recombination and cloning
3£®2 Construction of recombinant DNA molecules
3£®2£®1 Major classes of restriction endonucleases
3£®2£®2 Recognition sequences for type II restriction endonucleases
3£®2£®3 DNA ligase joins linear pieces of DNA
3£®2£®4 Sources of DNA for cloning
3£®3 Vectors for cloning
3£®3£®1 Genes coding for selectable markers are inserted into vectors during their construction
3£®3£®2 Plasmid DNA were the first cloning vector
3£®3£®3 Recombinant bacteriophages can serve as vectors
3£®3£®4 Cosmids and phagemids expand the repertoire of cloning vector
3£®4 Expression of recombinant genes
3£®4£®1 Expression vectors allow regulated and efficient expression of cloned genes
3£®4£®2 Expression systems
3£®5 Introducing recombinant DNA into host cells
3£®5£®1 Numerous host-specific techniques are used to introduce recombinant DNA molecules into living cells
3£®5£®2 Transient and stable transfection assays
3£®6 Constructing DNA libraries
3£®6£®1 Type of different libraries
3£®6£®2 Library screening and probes
3£®7 Sequencing of entire genomes
3£®7£®1 Genomic libraries contain the entire genome of an organism as a collection of recombinant DNA molecules
3£®7£®2 There are two approaches for sequencing large genomes
3£®8 Practical application of recombinant DNA technologies
3£®8£®1 Gene therapy
3£®8£®2 Delivering a gene into sufficient cells within a specific tissue and ensuring its subsequent long-term expression is a challenge
Chapter 4 Tools for Analyzing Gene Expression
4£®1 Introduction
4£®2 Gene isolation and detection
4£®2£®1 Agarose gel electrophoresis of DNA
4£®2£®2 Gradient centrifugation
4£®2£®3 Nucleic acid hybridization
4£®2£®4 Polymerase chain reaction
4£®2£®5 In vitro mutagenesis
4£®2£®6 DNA sequencing
4£®3 Analysis at the level of gene transcription£º RNA expression and localization
4£®3£®1 Reverse transcription PCR
4£®3£®2 Northern blotting
4£®3£®3 RNase protection assay (RPA)
4£®3£®4 In situ hybridization
4£®3£®5 DNA microarrays
4£®4 Analysis of the transcription rates
4£®4£®1 S1 nuclease protection and primer extension
4£®4£®2 Rapid amplification of cDNA ends £¬
4£®4£®3 Reporter genes
4£®4£®4 DNA footprinting
4£®4£®5 Electrophoretic mobility shift assay
4£®4£®6 Chromatin immunoprecipitation
4£®5 Analysis at the level of translation£º protein expression and localization
4£®5£®1 Western blotting
4£®5£®2 Enzyme-linke